转基因抗除草剂大豆ZH10-6转化体特异性定量检测体系的建立  

Establishing an specific quantitative detection system for transgenic herbicide-resistant soybean ZH10-6 transformant

作　　者：白蓝[1,2,3] 贾军伟 陈一帆[1,2,3] 章寅 潘爱虎 吴潇 BAI Lan;JIA Junwei;CHEN Yifan;ZHANG Yin;PAN Aihu;WU Xiao(Biotechnology Research Institute,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China;Shanghai Key Laboratory of Agricultural Genetics and Breeding,Crops Ecological Environment Security Inspection Center(Shanghai),Ministry of Agriculture and Rural Affairs,Shanghai 201106,China;Shanghai Professional Technology Service Platform of Agricultural Biosafety Evaluation and Testing,Shanghai 201106,China;Institute of Animal Husbandry and Veterinary Science,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China;Key Laboratory for Safety Assessment(Environment)of Agricultural Genetically Modified Organisms,Ministry of Agriculture and Rural Affairs,Shanghai 201106,China)

机构地区：[1]上海市农业科学院生物技术研究所,上海201106 [2]上海市农业遗传育种重点实验室,农业农村部农作物生态环境安全检验测试中心(上海),上海201106 [3]上海市农业生物安全评价与检测专业技术服务平台,上海201106 [4]上海市农业科学院畜牧兽医研究所,上海201106 [5]农业农村部农业转基因生物安全评价(环境)重点实验室,上海201106

出　　处：《上海农业学报》2024年第3期1-7,共7页Acta Agriculturae Shanghai

基　　金：上海市农业科学院卓越团队建设计划项目[沪农科卓(2022)016]。

摘　　要：以新型转基因抗除草剂大豆ZH10-6外源插入片段的旁侧序列为靶标,建立了转化体特异性实时荧光PCR和单通道微滴式数字PCR精准定量检测体系,并邀请8家农业农村部检测中心对检测方法的准确性进行验证。结果表明:该转化体特异性实时荧光定量检测方法具有良好的特异性、重复性及广泛的适用性,定量极限达到50 copies∕μL,检测极限达到5 copies∕μL。该检测方法有望作为检测标准用于转基因抗除草剂大豆ZH10-6的定量检测,为该大豆品系的商业化应用及转基因标识等监管提供技术支撑。A precise quantitative detection system for transformant specific real-time fluorescence PCR and single channel micro drop digital PCR was established using the flanking sequence of the exogenous insertion fragment of the novel genetically modified herbicide-resistant soybean ZH10-6 as the target.Eight testing centers of the Ministry of Agriculture and Rural Affairs were invited to verify the accuracy of the detection method.The results showed that the transformant specific real-time fluorescence quantitative detection method had good specificity,repeatability,and wide applicability,with a quantitative limit of 50 copies∕μL and a detection limit of 5 copies∕μL.This detection method was expected to serve as a detection standard for the quantitative detection of genetically modified herbicide-resistant soybean ZH10-6,providing technical support for the commercial application and regulation of genetically modified labeling of this soybean variety(line).

关 键 词：抗除草剂大豆ZH10-6 转化体特异性 定量检测 实时荧光PCR 微滴式数字PCR 

分 类 号：S336[农业科学—作物遗传育种]